title: Highly efficient biodiesel production by a whole-cell biocatalyst employing a system with high lipase expression in Aspergillus oryzae.
abstract: In the present study, a system with high lipase expression in Aspergillus oryzae was developed using an improved enolase promoter (P-enoA124) and the 5' untranslated region of a heat-shock protein (Hsp-UTR). P-enoA142 enhanced the transcriptional level of a heterologous lipase gene and Hsp-UTR improved its translational efficiency. Fusarium heterosporum lipase (FHL) was inserted into a pSENSU-FHL expression vector harboring P-enoA142 and Hsp-UTR and was transformed into an A. oryzae NS4 strain. Transformants possessing pSENSU-FHL in single (pSENSU-FHL#1) and double copies (pSENSU-FHL#2) were selected to evaluate the lipase activity of the whole-cell biocatalyst. The two strains, pSENSU-FHL#1 and #2, showed excellent lipase activity in hydrolysis compared with the strain transformed with conventional expression vector pNAN8142-FHL. Furthermore, by using pSENSU-FHL#2, methanolysis could proceed much more effectively without deactivation, which allowed a swift addition of methanol to the reaction mixture, thereby reducing reaction time.
authors.affiliation: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkoudaicho, Nada-ku, Kobe 657-8501, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Takaya
authors.forename: Risa
authors.initials: R
authors.lastname: Koda
authors.forename: Daisuke
authors.initials: D
authors.lastname: Adachi
authors.forename: Kazunori
authors.initials: K
authors.lastname: Nakashima
authors.forename: Junpei
authors.initials: J
authors.lastname: Wada
authors.forename: Takayuki
authors.initials: T
authors.lastname: Bogaki
authors.forename: Chiaki
authors.initials: C
authors.lastname: Ogino
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kondo
mesh_terms: D001236:Aspergillus oryzae / Q000235:genetics* / Q000378:metabolism; D055162:Biocatalysis; D056804:Biofuels / Q000032:analysis* / Q000382:microbiology; D001709:Biotechnology / Q000295:instrumentation / Q000379:methods*; D018914:Cells, Immobilized / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005670:Fusarium / Q000201:enzymology; D015870:Gene Expression; D008049:Lipase / Q000235:genetics / Q000378:metabolism*; D000432:Methanol / Q000378:metabolism*
chemical_list: D056804:Biofuels; D005656:Fungal Proteins; D008049:Lipase; D000432:Methanol